Related Experiment Video
Updated: Jun 17, 2026
High-throughput Screening of Carbohydrate-degrading Enzymes Using Novel Insoluble Chromogenic Substrate Assay Kits
Published on: September 20, 2016
Fluorogenic substrates for the screening assay of transketolase through beta-elimination of
F Charmantray1, B Légeret, V Hélaine
1Clermont Université, Université Blaise Pascal, Laboratoire SEESIB, BP 10448, F-63000 Clermont-Ferrand, France. Franck.CHARMANTRAY@univ-bpclermont.fr
Abstract:
5-O-Coumarinyl-d-xylulose was studied as a fluorogenic substrate for the stereospecific assay of transketolase enzyme. Enzymatic C2-C3 cleavage released an alpha-hydroxyl, beta-coumarinyl substituted aldehyde. Although the subsequent beta-elimination step was rate limiting under chemical or enzymatic catalysis, we detected a TK activity as low as 0.7mIU. To improve the fluorescence signal release, kinetic and product distribution analyses of this reaction were performed by LC/UV/MS coupling.
